Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskålea

Franklyn G. Prendergast, Kenneth G. Mann

Research output: Contribution to journalArticle

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Abstract

The calcium-sensitive photoprotein aequorin and the green fluorescent protein were isolated and purified from Aequorea forskålea. Purified aequorin shows electrophoretic microheterogeneity but appears as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino-terminal sequence determination by the automated Edman technique revealed a single NH2-terminal sequence: Val-Lys-Leu-(Thr)-Pro-Asp- Phe-Asn-Asn-Pro-(-)-Trp-Ile-Gly-Arg-His-aequorin exists as a single polypeptide chain. Apparent molecular weights were determined by sedimentation equilibrium, urea-sodium dodecyl sulfate/polyacrylamide gel electrophoresis, gel filtration of native protein, and gel filtration of denatured protein in 6 M guanidine hydrochloride; all methods suggest an apparent molecular weight of 19 500 ± 2000 for aequorin. Under some conditions the protein dimerizes by disulfide bond formation. Aequorin has a sedimentation coefficient of 2.31 S and a Stokes radius of ∼19 Å; the extinction coefficient (E1cm 1%) was calculated to be 27.1. Amino acid analysis revealed a slight preponderance of acidic residues; no carbohydrate moieties were found. Aequorin contains at least one free sulfhydryl group, chemical modification of which results in irreversible loss of luminescent activity. The apparent molecular weight of the green fluorescent protein is 30 000 when determined by urea-sodium dodecyl sulfate/polyacrylamide gel electrophoresis and gel filtration of denatured protein in 6 M guanidine hydrochloride. Amino acid analysis revealed an absence of tryptophan.

Original languageEnglish (US)
Pages (from-to)3448-3453
Number of pages6
JournalBiochemistry
Volume17
Issue number17
StatePublished - 1978

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Aequorin
Green Fluorescent Proteins
Chemical properties
Physical properties
Electrophoresis
Sodium Dodecyl Sulfate
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Molecular Weight
Gels
Molecular weight
Guanidine
Sedimentation
Urea
Proteins
Luminescent Proteins
Amino Acids
Chemical modification
Tryptophan
Disulfides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskålea. / Prendergast, Franklyn G.; Mann, Kenneth G.

In: Biochemistry, Vol. 17, No. 17, 1978, p. 3448-3453.

Research output: Contribution to journalArticle

Prendergast, Franklyn G. ; Mann, Kenneth G. / Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskålea. In: Biochemistry. 1978 ; Vol. 17, No. 17. pp. 3448-3453.
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